茶树花发育相关的一个钙依赖蛋白激酶基因的克隆与表达分析  

Molecular Cloning and Expression of a Flowering-related Calcium-dependent Protein Kinase Gene in Tea Plant [Camellia sinensis (L.)O.Kuntze]

作　　者：陈暄[1] 汤茶琴[1,2] 邹中伟[1] 杨亦扬[1] 房婉萍[1] 王玉花 黎星辉[1] 

机构地区：[1]南京农业大学茶叶科学研究所,江苏南京210095 [2]无锡市茶叶品种研究所,江苏无锡214063

出　　处：《茶叶科学》2009年第1期47-52,共6页Journal of Tea Science

基　　金：教育部高校博士点基金(20060307024);南京农业大学青年科技创新基金(KJ06007);江苏省三项工程(sx2007g17、sx2008g10);江苏省科技攻关计划(BE2007301、BE2008320-2);国家自然科学基金(30800884)资助

摘　　要：应用cDNA-AFLP(Amplified Fragment Length Polymorphism)技术分析了结实率差异显著的龙井43和大叶乌龙两个茶树品种在花蕾发育过程中基因表达的差异。从获得的差异图谱中,克隆得到一个与茶树花发育相关的钙依赖蛋白激酶基因片段,然后用RCAE方法扩增获得其cDNA全长序列,命名为茶树TCK(Camellia sinensis calcium-dependent protein kinase)基因,GenBank登录号EU732607。该基因cDNA序列全长2281bp,编码760个氨基酸。用RT-PCR方法进一步研究该基因的功能,检测其表达特异性,结果表明该基因只在茶树花蕾发育后期特异表达,在叶、花蕾发育早期均无表达,提示TCK基因可能在茶树花发育过程中发挥重要作用。To have an insight into the comprehensive molecular characteristics relating to the flower development mechanisms of tea plant [Camellia sinensis （L.）O.Kuntze], cDNA-AFLP （Amplified Fragment Length Polymorphism） was performed within two tea plant cultivars, Longjin43 and Daye Wulong. A cDNA fragment encoding a calcium-dependent protein kinase gene was isolated and identified. The complete cDNA sequence was cloned by RACE （rapid amplification of cDNA ends）, named TCK（Camellia sinensis calcium-dependent protein kinase）gene. GenBank accession number is EU732607. To identify differential expression of TCK gene, RT-PCR experiments were performed in six samples including leaves, small flower buds and big flower buds of Longjing 43 and Daye Wulong. Results showed that the expect band exists in the big flower buds, no band in leaves and small flower buds of these two cultivars. It hinted that TCK might play a key role during flower development of tea plant.

关 键 词：茶树 花发育 钙依赖蛋白激酶基因 克隆 特异表达 

分 类 号：S571.1[农业科学—茶叶生产加工] Q523[农业科学—作物学]